FAST SOLID SUPPORT DETECTION OF HUMAN PAPILLOMAVIRUS IN IN-VITRO AMPLIFIED DNA USING A DNP ANTI DNP MONOCLONAL-ANTIBODY COUPLE

In some anogenital lesions the detection of certain types of human pap illoma virus, especially oncogenic types, is of interest. In a first s tep during a prospective study, we compared two methods for the detect ion of human papillomavirus (HPV) DNA in clinical samples: Southern bl otting followed by hybridization with a cloned radioactive genomic pro be and a classical polymerase chain reaction (PCR) followed by hybridi zation with a (32)p- labelled oligonucleotide probe. 118 biopsies and swabs were examined for HPV 6/11, 16, 18 and 33, 67 positive reactions were found by both methods, 5 positives only by PCR and 2 positives o nly by Southern blot for unidentified HPV. Patients with anogenital co ndylomas, dysplasias and carcinomas or asymptomatic patients were stud ied. Most high grade (II and III) dysplasias were associated with HPV 16 and HPV 18. Condylomata lesions and low grade dysplasia (grade I) w ere associated mostly with HPV 6/11, mixed type of HPV, less frequentl y with HPV 16 or HPV 18. As a second step a nested PCR coupled to soli d support detection method was used as described by Sauvaigo et al. (1 990) Nucleic Acids Res. 18, 3175-3183) to study a panel of 30 previous ly qualified different HPV DNA extracts. In this procedure the second round of PCR amplification involves biotinylated and dinitrophenylated labelled primers,allowing the capture of PCR amplified HPV DNA sequen ces on streptavidin coated tubes and its revelation. We describe an im provement of HPV DNA detection by means of single-step immunoenzymatic revelation involving anti-DNP monoclonal antibodies conjugated to hor seradish peroxidase enzyme. A perfect correlation with the previous re sults was obtained. This solid support method allows a faster and easi er HPV typing compared to methods using membrane transfer.